Amelioration of beta654-thalassemia in mouse model with the knockdown of aberrantly spliced beta-globin mRNA.
Large amounts of aberrantly spliced mRNA from the beta(654) allele was present in erythroid cells, which might impair the erythropoiesis. A therapeutic strategy for beta-thalassemia was explored by knocking down the aberrantly spliced mRNA of beta-globin. Lentiviral vector with siRNA fragment targets on the specific portion of beta(654)-globin aberrantly spliced pre-mRNA was constructed. In HeLa beta(654) cells, the siRNA vector could reduce approximately 60% of aberrantly spliced mRNA, which was assessed by RT-PCR and qRT-PCR. Furthermore, a disease model of beta(654) thalassemia mice with lentiviral-mediated siRNA was produced by subzonal injection (named Hbetai-Hbb(th-4)/Hbb(+) transgenic mice). Our results showed that the hemotological parameters were improved in Hbetai-Hbb(th-4)/Hbb(+) transgenic mice. This study provides a potential way for beta(654)-thalassemia therapy by knocking down the aberrantly spliced beta-globin mRNA, whilst supporting that the aberrantly spliced beta-globin mRNA may aggravate the disease.
Xie
Shuyang
S
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200040, China.
Li
Wei
W
Ren
Zhaorui
Z
Zhang
Jingzhi
J
Guo
Xinbin
X
Wang
Shu
S
Huang
Shuzhen
S
Zeng
Fanyi
F
Zeng
Yi-Tao
YT
D017398:Alternative Splicing / Q000235:genetics*; D000818:Animals; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D010641:Phenotype; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000235:genetics* / Q000473:pathology / Q000628:therapy*
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D055544:beta-Globins